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Identification and Functional Expression of a Δ9‐Fatty Acid Desaturase from Psychrobacter urativorans in Escherichia coli
Author(s) -
Li Yan,
Dietrich Matthias,
Schmid Rolf D.,
He Bingfang,
Ouyang Pingkai,
Urlacher Vlada B.
Publication year - 2008
Publication title -
lipids
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.601
H-Index - 120
eISSN - 1558-9307
pISSN - 0024-4201
DOI - 10.1007/s11745-007-3150-5
Subject(s) - biochemistry , escherichia coli , fatty acid , biology , palmitoleic acid , fatty acid desaturase , palmitic acid , fatty acid synthesis , gene , chemistry , polyunsaturated fatty acid
The Δ9‐fatty acid desaturase is a key enzyme in the synthesis of unsaturated fatty acids. The fatty acid composition of membrane phospholipids in Psychrobacter urativorans is characterized by a high degree of desaturation at Δ9 position. Based on CODEHOP‐mediated PCR strategy, a novel gene designated as PuFAD9 , putatively encoding a Δ9‐fatty acid desaturase ( Pu FAD9), was isolated from P. urativorans . The gene consists of 1,455 bp and codes for 484 amino acids. Analysis of the amino acid sequence reveals three histidine clusters and a hydropathy profile, typical for membrane‐bound desaturases. Activity of the Pu FAD9 protein, recombinantly expressed in Escherichia coli was confirmed by GC‐MS analysis of the cellular fatty acid composition. It was found that the ratio between palmitoleic and palmitic acid in E. coli cells heterologously expressing the PuFAD9 gene was significantly affected by IPTG induction and the growth temperature.